MRM Proteomics and Agilent Technologies Introduce First Commercial Kits for Quality Control of MRM-Based Quantitative Plasma Proteomic Workflows

ANTA CLARA, Calif., and VANCOUVER, B.C., Oct. 9, 2013 – Agilent Technologies Inc. (NYSE: A) and MRM Proteomics Inc. today announced the launch of MRM Proteomics’ PeptiQuant MRM Assay Kits for quality control in  quantitative plasma proteomic analyses based on multiple-reaction monitoring.

The two new kits are designed for use on Agilent’s latest-generation standard-flow UHPLC-QQQ MS/MS platform, which consists of the 1290 Infinity UHPLC system and the 6490 triple quadrupole mass spectrometer platform.

MRM Proteomics developed the kits to assist in the inter- and intra-laboratory quality control of MRM experiments in conjunction with stable isotope-labeled standards:

  • The PeptiQuant LC-MS Platform Performance Kit tests the effectiveness of the LC-MS platform.
  • The PeptiQuant Workflow Performance Kit evaluates the performance of an entire LC-MRM-MS analytical workflow in human plasma from denaturation through digestion to detection.

“Mass spectrometry is becoming increasingly important in biomedical and biological research, especially with the launch of large-scale initiatives such as the Human Proteome Project, which depends on mass spectrometry as one of the main technology platforms,” said Andrew Munk, CEO of MRM Proteomics. “The mass spec and proteomics communities currently lack standardized protocols and reagents to ensure high-quality data that can be reproduced by laboratories across the world using different mass-spec technologies. Our PeptiQuant™ targeted proteomics Platform and Workflow performance kits will ensure that high-quality data can be reproduced.”

“Application of QQQ mass spectrometry to protein quantitation is growing rapidly, and the adoption of this technique will be dependent on robust protocols that can be used in multiple laboratories,” said Steven Fischer, director of omics applications for Agilent. “Agilent is pleased to be working with MRM Proteomics to address this need. PeptiQuant kits from MRM Proteomics, combined with Agilent’s UHPLC-QQQ MS/MS platform, can be used to ensure high-quality protein quantification data is being produced in labs around the world.”

 

About MRM Proteomics

MRM Proteomics Inc. is a leader in providing advanced protein quantitation, analytical proteomic services and reagents to the pharmaceutical, biotechnology and diagnostics industries in key areas such as biomarker discovery/validation and clinical diagnostics development. Information about MRM Proteomics is available at www.mrmproteomics.com.

 

About Agilent Technologies

Agilent Technologies Inc. (NYSE:A) is the world’s premier measurement company and a technology leader in chemical analysis, life sciences, diagnostics, electronics and communications. The company’s 20,500 employees serve customers in more than 100 countries. Agilent had revenues of $6.9 billion in fiscal 2012. Information about Agilent is available at www.agilent.com.

 

EDITORIAL CONTACTS:

Agilent Technologies

Eric Endicott

+ 1 408 553 2005

eric_endicott@agilent.com

 

MRM Proteomics Inc.

Andrew Munk, CEO

+1 604 800 2296

munk@mrmproteomics.com

MRM Proteomics and PROOF Centre Collaborate with the Jain Foundation to Identify Blood-Based Biomarkers of LGMD2B/Miyoshi Muscular Dystrophy

Vancouver, BC and Seattle, WA – The Centre of Excellence for the Prevention of Organ Failure (PROOF Centre) and MRM Proteomics Inc. will use their biomarker expertise in collaboration with the Jain Foundation, Inc. to identify blood-based biomarkers of disease in individuals with a rare form of muscular dystrophy, called Limb‐girdle muscular dystrophy 2B (LGMD2B) or Miyoshi Myopathy (MM). Since no effective treatment exists for LGMD2B/ MM, this study is an essential step towards developing therapies for patients living with these debilitating muscle diseases.

LGMD2B/MM are both caused by mutations in the gene that encodes the protein dysferlin, which is thought to play a role in skeletal muscle repair collectively called “dysferlinopathies.”

The Jain Foundation chose to collaborate with MRM Proteomics and the PROOF Centre because of their excellent track record of discovering molecular biomarkers in blood and developing them into tests that aid in the diagnosis and treatment of other diseases.

“The Jain Foundation was impressed by the success PROOF Centre has had in moving biomarker programs from concept to the clinic, particularly with their recent clinical progress in finding a biomarker for rejection of a transplanted heart. We believe that pairing MRM Proteomics’ cutting edge expertise in mass spectrometry with PROOF’s unique clinical biomarker expertise, offers the greatest chance of identifying a biomarker to move our clinical program forward.” said Plavi MIttal, President and CEO, Jain Foundation.

The study will compare blood samples from individuals with LGMD2B/MM with samples from healthy, age and gender-matched controls. Differences in proteins and nucleic acids in the blood that reflect differences in muscle function will be tested and validated as possible biomarkers. These biomarkers will then be used to help track disease progression and the efficacy of potential treatments in future clinical trials.

 

About MRM Proteomics

MRM Proteomics Inc. is a leader in providing advanced mass spectrometry based quantitative proteomics services and reagents to the pharmaceutical, biotechnology and diagnostics industries in key areas such as biomarker discovery/validation and clinical diagnostics development. www.mrmproteomics.com.

 

About the Proof Centre of Excellence

The Centre of Excellence for the Prevention of Organ Failure (PROOF Centre) develops blood tests to diagnose, treat and manage heart, lung and kidney disease. The PROOF Centre is a cross-disciplinary engine of partners representing academia, health care, government, industry, patients and the public, to improve heart, lung and kidney health and reduce the enormous socioeconomic burden of organ disease. The centre’s service arm provides expertise in computation, “-omic” science and biomarker workflow to organizations seeking to discover, develop and implement blood-based molecular biomarkers in a clinical setting. The PROOF Centre is a not-for-profit society initially established from the Networks of Centres of Excellence Secretariat under the Centre of Excellence for Commercialization and Research (NCE CECR) Program, and is co-hosted by the University of British Columbia and Providence Health Care in Vancouver, British Columbia, Canada. www.proofcentre.ca.

 

About Jain Foundation

The Jain Foundation is a non-profit organization with a mission to identify a cure or therapies for LGMD2B/MM. The foundation is privately funded and does not solicit funding from patients or other sources. The foundation’s activities include providing financial support to scientific researchers, identifying promising drug candidates, funding clinical trials and studies, educating patients and helping patients obtain genetic diagnoses. www.jain-foundation.org.

 

Contacts:

Jerrie Lynn Morrison

Communications Coordinator

PROOF Centre of Excellence

Phone: 604-682-2344 ext 63729

Email: jerrielynn.morrison@hli.ubc.ca

 

Andrew Munk

Chief Executive Officer

MRM Proteomics Inc.

Phone: 604-800-2296

Email: munk@mrmproteomics.com

 

Sarah Shira

Office and Program Manager

Jain Foundation

Phone: 425-882-1492

Email: admin@jain-foundation.org

 

MRM Proteomics and PROOF Centre of Excellence Partner to Bring New Diagnostic Tests to Clinic

Two British Columbian biotech organizations collaborate to optimize development of blood tests for patients with chronic organ disease

Vancouver, BC – The Centre of Excellence for the Prevention of Organ Failure (PROOF Centre) and MRM Proteomics are collaborating to bring new diagnostic blood tests which will help clinicians manage patients with heart, lung and kidney disease more effectively.

The PROOF Centre – a not-for-profit organization focused on developing blood tests for chronic organ disease – and MRM Proteomics, a contract research organization, are integrating their expertise to create an end-to-end solution that uses protein signatures in the blood to predict the onset and progression of heart, lung, and kidney disease.

The PROOF Centre’s award-winning computational pipeline will be applied to guide the experimental design, and data quality control and analysis needed for the identification of protein signatures that can be used with other molecular signals to indicate heart, lung, and kidney disease. With its highly-skilled computation team of database specialists, clinical statisticians, bioinformaticians and computer and systems scientists, the PROOF Centre will support the analysis process from front-end experimental design to downstream statistical and biological analysis.

“This is a great way to begin to commercialize our strengths in the internationally-recognized computation capabilities that we’ve built up over the past few years,” says Dr. Bruce McManus, Director of the PROOF Centre. “Through this collaboration, MRM Proteomics’ expertise and leading technology will help us bring these tests to the clinic faster, ultimately helping improve patient care and lower heath costs.”

MRM Proteomics specializes in the absolute quantification of proteins in complex biological fluids such as blood and urine using Multiple Reaction Monitoring -Mass Spectrometry (MRM-MS). Their PeptiQuant™ MRM assays allow for the absolute quantification of hundreds of proteins at a time with unparalleled specificity and sensitivity, from a single drop of blood. These services lead to faster results for lower costs and sample volumes than other protein biomarker quantification methods.

“Our organizations’ offerings are clearly complementary in the field of protein biomarker discovery and validation.” said Andrew Munk, President and CEO of MRM Proteomics. “The discovery, validation and implementation of biosignatures are complex processes. Partnering with the PROOF Centre will provide our customers with access to the PROOF Centre’s internationally recognised computational tools and capabilities to optimize the design and analysis of their mass spec based proteomic biomarker studies.”

About MRM Proteomics:

 MRM Proteomics is a leader in providing advanced protein quantitation and proteomic services to the pharmaceutical, biotechnology and diagnostics industries in key areas such as biomarker discovery/validation, clinical studies, diagnostics and toxicology. Information about MRM Proteomics is available at www.mrmproteomics.com.

About the PROOF Centre:

The Centre of Excellence for the Prevention of Organ Failure (PROOF Centre) accelerates the development of new blood tests to improve all aspects of care for patients with heart, lung and kidney disease. The PROOF Centre is a cross-disciplinary engine of partners representing academia, health care, government, industry, patients and the public, focused on improving health and reducing the enormous socioeconomic burden of heart, lung and kidney disease. The PROOF Centre is a not-for-profit society established in March 2008 by competitive funding from the Networks of Centres of Excellence Secretariat under the Centre of Excellence for Commercialization and Research (NCE CECR) Program. The PROOF Centre is co-hosted by the University of British Columbia and Providence Health Care in Vancouver, British Columbia, Canada.

MRM Proteomics, Agilent Technologies Sign Co-marketing Agreement to Further Quantitative Proteomics Solutions

SANTA CLARA, Calif. and VANCOUVER, B.C., Sept. 13, 2012 – Agilent Technologies Inc. (NYSE: A) today announced that it has entered into a comarketing agreement with MRM Proteomics Inc., a leading provider of advanced protein quantitation, biomarker and proteomic services to the pharmaceutical, biotechnology and diagnostics industries. The agreement will allow both organizations to offer a more complete solution, including kits, hardware and software for proteomics.

Proteomics is the study of the structure and function of proteins and how they interact within a complex biological system. Agilent’s integrated proteomics workflow provides the highest analytical performance with unprecedented plug-and-play flexibility. Interchangeable workflows simplify setups and let researchers quickly switch between different methodologies. Offering the industry’s highest sensitivity and highest throughput quantitation, the Agilent 6490 Triple Quadrupole MS/MS provides targeted MRM peptide quantitation at attomole detection levels.

          “MRM Proteomics, is recognized as a pioneer and leading company in the development of mass spec based methods for protein quantification using multiple reaction monitoring,” said Steve Fischer, Agilent marketing manager, Metabolomics and Proteomics. “The agreement is a further reflection of Agilent’s continued commitment and investment in quantitative proteomics.”

MRM Proteomics specializes in the highly multiplexed absolute quantitation of proteins in complex biological samples such as blood, cerebrospinal fluid and urine through MRM-MS using isotopically labeled internal standards. MRM Proteomics’ workflows utilize minimal sample volumes (20 ul) and are capable of unparalleled specificity and sensitivity, without the need for depletion of high-abundance proteins.

“MRM Proteomics is pleased to enter into this comarketing arrangement with Agilent Technologies; our companies’ offerings are clearly complementary in the field of targeted quantitative proteomics,” said Andrew Munk, chief executive officer of MRM Proteomics. “Mass spectrometry is a rapidly growing platform driving basic proteomics research, biomarker discovery and drug development. Agilent’s status as an innovator in mass spec instrumentation combined with its extensive global network, make Agilent a great partner for MRM Proteomics.

About MRM Proteomics

MRM Proteomics is a leader in providing advanced protein quantitation and proteomic services to the pharmaceutical, biotechnology and diagnostics industries in key areas such as biomarker discovery/validation, clinical studies, diagnostics and toxicology. Information about MRM Proteomics is available at www.mrmproteomics.com.

About Agilent Technologies

Agilent Technologies Inc. (NYSE: A) is the world’s premier measurement company and a technology leader in chemical analysis, life sciences, diagnostics, electronics and communications. The company’s 20,000 employees serve customers in more than 100 countries. Agilent had net revenues of $6.6 billion in fiscal 2011. Information about Agilent is available at www.agilent.com.

Contacts:
Andrew Munk
President and CEO
+1 (604) 800-2296
munk@mrmproteomics.com

Multiplexed quantification of 63 proteins in human urine by multiple reaction monitoring (MRM)-based mass spectrometry (MRM-MS)

Urine samples from 156 patients with hernia, bladder cancer, or urinary tract infection/hematuria were subjected to multiplexed LC-MRM/MS to determine the absolute concentrations of the 63 proteins in urine.  Stable isotope-labeled standard proteotypic peptides were used as internal standards for the 63 targeted proteins. All peptides were synthesized in-house for the highest purity and quality control.

Twelve proteins showed higher concentrations in the bladder cancer group than in the hernia and the urinary tract infection/hematuria groups, and thus represent potential urinary biomarkers for detection of bladder cancer.

The multiplexed MRM-MS data was used to generate a six-peptide marker panel. This six-peptide panel could discriminate bladder cancer subjects from non-cancerous subjects with an AUC of 0.814, with a 76.3% positive predictive value, and a 77.5% negative predictive value.

Like to learn more about highly multiplexed biomarker discovery and/or biomarker validation in urine, plasma or cerebrospinal fluid (CSF) using MRM mass spec? Find out more about our MRM Assays at MRM Proteomics here: www.mrmproteomics.com

A MRM-based Multiplexed Quantitation of Putative Cardiovascular Disease Biomarkers in Human Plasma

We have developed a highly-multiplexed multpile reaction monitoring (MRM-MS) MRM-based assay for determination of cardiovascular disease (CVD) status and disease classification.

The assay uses stable-isotope-labeled peptide standards (Internal Standards) for the quantitation of putative biomarkers of CVD in tryptic digests of whole plasma in a 30-min assay. The assay allows for absolute quantification of proteins in plasma.

Eighty-five analyses of the same sample showed no loss of sensitivity (<20% CV) and no loss of retention time accuracy (<0.5% CV for all peptides). The maximum linear dynamic range of the MRM assays ranged from 103-10for the assays. Excellent linear responses (r >0.98) were obtained for the peptide targets with attomole level limits of quantitation (<20% CV and accuracy 80-120%). The assay allows for the absolute quantiation of proteins down to 2 nanograms of protein per microliter of plasma (ng /ml) in undepleted unfractionated plasma.

Although depletion strategies have been used for plasma biomarker discovery studies, the ideal analytical workflow for biomarker validation or clinical assays should not require depletion as these procedures also remove diagnostically-useful high-abundance proteins (such as low density lipoprotein), as well as adding additional cost, complexity, and time to the analysis. Moreover, depletion methods may compromise analytical reproducibility. For this reason no depletion is used in any of our MRM-MS quantitative assays.

The assay is robust, sensitive (ng/ml), and has high-throughput capabilities through short analysis time and complete automated sample preparation. It is therefore well suited for CVD biomarker validation and discovery in plasma.

Interested in this MRM assay? Find out more about MRM Proteomics’ PeptiQuantTM MRM Assays here: http://www.mrmproteomics.com

What are the advantages of Multiple Reaction Monitoring over Immunoassays for protein quantification?

What are the advantages of Multiple Reaction Monitoring over Immunoassays for protein quantification?

PeptiQuant™ MRM Assays – Advantages

PeptiQuant Assays have several significant advantages over other protein quantification (biomarker quantification) methods, including HPLC and antibody based ELISA assays.

  • Short assay development time – 10 to 16 weeks.
  • No antibody development costs 
  • Highly Multiplexed (10s to 100s)
  • Absolute quantification 
  • Absolute molecular specificity
  • Same analysis can be applied in preclinical & clinical testing
  • Minimal sample volumes (10-50ul)

You can learn more about MRM assays here

PeptiQuant™ MRM Assays – Wide number of applications in drug discovery and clinical development including:

  • Biomarker discovery, development and verification
  • Probing signaling pathways
  • Drug toxicity assessments in serum samples (Toxicoproteomics)
  • Patient selection markers & clinical trial monitoring
  • Pharmacokinetic (pK) studies

 Multiple Reaction Monitoring (MRM) Mass Spectroscopy

Multiple Reaction Monitoring (MRM) is highly selective (targeted), allowing researchers to fine tune an instrument to specifically look for peptides, or protein fragments, of interest. This approach allows for greater specificity, sensitivity, speed and quantitation of an analyte of interest (eg. biomarker candidate).

For the application of biomarker discovery, verification and validation — in which absolute quantification of a single protein or a panel of candidate proteins in a large number of samples is required — a high throughput and multiplexing analysis is essential. The multiplex capability of mass spectrometry based platforms to quantitatively detect multiple candidate peptides in a complex sample is substantial and has been well demonstrated in different sample types and with different mass spectrometric protocols. It is practically feasible that more than 100 candidates could be simultaneously detected with absolute quantification in a single mass spectrometric measurement.

Immunoassays Have Problems

  • Anti-reagent antibodies, Anti-analyte autoantibodies
  • What actually forms/prevents the immunoassay sandwich is not directly visualized (false positive/negative results)
  • Cross-reactivity
  • Limited multiplexing (12 analytes/analysis)
  • Poor interplatform concordance
  • Immunoassay-grade Abs do not exist for vast majority of proteins
  • Expensive & time consuming to design and implement

Enzyme Linked Immunosorbent Assay (ELISA)

Enzyme linked immunosorbent assay (ELISA) based assays are presently the “Gold Standard” with reference to the detection of circulating blood biomarkers and are currently the major method for the quantification of proteins. This is commonly accomplished by sandwich ELISA using one antibody for capture of the circulating protein and another antibody to visibly detect antibody-protein binding, allowing for confirmation of protein presence and, in the case of ELISA, quantity.

However, the limiting factors for the ELISA as a technique for plasma biomarker verification are the restricted possibility to multiplex assays and the availability of antibodies for novel candidate proteins, combined with the lengthy and expensive development of new assays. ELISA based techniques require antibodies specific to the protein(s) that must be detected or quantified. This poses a problem as antibodies are often unavailable for many potential biomarkers which are novel proteins of unknown function. Even if antibodies are available for the proteins in question, throughput is limited as these techniques must be performed on a protein by protein basis and true multiplexing is limited.